A20 inhibits lipopolysaccharide-induced inflammation in enterocytes

• 作者: Cui-Fang Zheng Jie-Ru Shi Sheng-Nan Wang Ying Huang
• 来源期刊: 世界胃肠药理与治疗学杂志：英文版（电子版）
• 学科: 医学
• 关键词: A20 (TNFAIP3 ) LIPOPOLYSACCHARIDE 原子 factor-&kappa B 煽动性的肠疾病
• 年，卷（期）: 2016,（4）
• 页码范围: 540-549
• 页数: 10页
• 分类号: R574

摘要

AIM To examine the role of A20 in the regulation of intestinal epithelial cells(IECs) inflammation.METHODS Using gene transfection,both stable overexpression and knockdown A20-expressed HT-29 cell lines were established.Accordingly,the cells were divided into the following groups:The control group,the A20 overexpression group,the A20 knockdown group and the respective controls.A20 was stimulated with lipo-polysaccharide(LPS) in a dose- and time-dependent manner and was detected using western blotting and real-time polymerase chain reaction(PCR) analyses.Immunofluorescence and western blotting analyses were performed to investigate the role of A20 in the regulation of nuclear factor(NF)-κB activation and translocation into the nucleus.ELISA and real-time PCR were performed to examine A20 in regulating the release of the following inflammatory cytokines:Tumor necrosis factor(TNF)-a,interleukin(IL)-1b,IL-6 and IL-8.RESULTS The expression of A20 in IECs was inducible.When intestinal epithelial cells were subjected to the stimulation of LPS,the expression of A20 was increased,and the expression of A20 was induced in a dose-and timedependent manner.The expression of A20 was very low in HT-29 cells without LPS stimulation but rapidly increased and was maintained at a high level 2-4 h after stimulationwith LPS.These levels gradually declined with a change in time-course,and the expression of A20 increased with increasing LPS stimulation.Western blotting and immunofluorescence revealed that overexpression of A20 can inhibit NF-κB activation and its translocation to the nucleus.The overexpression of A20 can reduce the levels of proinflammatory cytokines involved in the pathophysiology of inflammatory bowel disease.There was no significant difference in the expression of IL-8 mR NA in the control group,A20 overexpression group or A20 knockdown group without LPS stimulation(P > 0.05);however,while after 2 h,4 h and 8 h stimulation with LPS,the expression of IL-8 in the A20 overexpression group was lower than the control gr